ABSTRACT

Crude oil processing produces toxic byproducts, such as benzene, toluene, and xylenes (BTX) capable of contaminating groundwater and soil leading to potentially serious health risks for both humans and wildlife. Thus, the effective removal of these hazardous organic compounds is imperative towards protecting natural resources. While it is possible to manually clean contaminated sites, such efforts are costly and time consuming. Our project seeks to degrade the toluene-based contaminants into valuable aromatic aldehydes by expressing the xyl ortho pathway from Pseudomonas putida in E. coli. As a continuation of our 2016 project, the 2017 UT-Knoxville iGEM team aims to use synthetic biology and metabolic engineering techniques to further develop our bioremediation strain. First, regulatory elements sensitive to the presence of aromatic hydrocarbons are engineered to fine-tune protein production. Second, overexpression of broadly specific efflux pumps intend to increase organic solvent tolerance and alleviate toxic effects. And third, exploring enzymatic homologs can expand our product library. Our project aims to increase production titers and develop a more robust microorganism suitable for manufacturing and bioremediation needs.